Dual chamber reagent mixing container

ABSTRACT

Described is an automated reagent mixing container for separately storing and automatically mixing together at least two stored reagent components.

CROSS REFERENCE TO RELATED APPLICATION

This application is a divisional application of co-pending U.S.application Ser. No. 15/234,138 filed on Aug. 11, 2016. The entirecontents of which are incorporated herein in its entirety by referencefor all purposes.

TECHNICAL FIELD OF THE INVENTION

The present invention is related to the field of automated clinicaldiagnostics, more specifically containers for storing and mixingcomponents of reagents used in diagnostic tests conducted in anautomated clinical analyzer, methods of use, and methods of makingthereof.

BACKGROUND

In many in-vitro diagnostic (IVD) testing procedures there is a need toprepare reagents by reconstituting and/or mixing multiple necessarycomponents, some in a liquid and others in a powder format. Currently,reconstituting or mixing reagents for an IVD procedure is achieved bymanual operation. For instance, in a diagnostic testing lab, beforeusing the reagents for sample testing, typically body fluids such aswhole blood, plasma, serum, urine, cerebrospinal fluid and so on,medical workers collect different reagent components from separatevials/bottles, use a pipette to pipette diluents with a certain volumeto a vial of powder reagents or to a vial with concentrated liquidreagents, wait for certain time for reconstitution, and finally mix thereagents manually by shaking, stirring, or rotating, for example. Thismanual process reduces the speed in which a diagnostic test takes place,increases the risk of human error and operator contamination withpotentially toxic chemicals, and raises the cost of packaging.

Dual-chamber syringe systems developed for drug preparation, provide asolution for mixing drug components such as a lyophilized activecomponent and a diluent like water or saline, by packaging two drugcomponents in a single device such as a dual barrel syringe therebypartially automating the drug reconstitution process. However, thesyringe device is disadvantaged by a complex manufacturing and assemblyprocess. The cost of the manufacturing and assembly is not a factor fordrug manufacturers in view of the price of the drug to the consumer. Butsuch devices are not practical for IVD test applications, where thepackaging cost for IVD test applications must be controlled to be verylow because of the low cost of the test to the consumer.

In addition, the volume inside a syringe is relatively low, compared toreagent volume in IVD testing, potentially a liter or more, such as atest run by an automated clinical analyzer, to achieve effectivereconstitution and mixing. In addition to the need to reconstitutereagents for IVD applications, the contents of current reagent vialsused in diagnostic instruments such as automated clinical analyzers, aresubject to evaporation when on-board the instrument. Such evaporationcompromises reagent stability, alters reagent concentration, and isgenerally wasteful of reagents, increasing the cost for the user.

In order to improve the current reagent preparation process and usageefficiency for IVD applications in a clinical analyzer, a reagentpackaging solution with multi-components storage, automatedreconstitution and mixing, and evaporation prevention is needed.

SUMMARY OF THE INVENTION

The present invention relates to the field of clinical diagnosis,diagnostic assays in particular, and more specifically containers andmethods for storing and automated mixing of reagent components for usein a diagnostic assay in an automated clinical analyzer.

In one aspect, the invention described herein is related to a reagentmixing container having two integrated chambers: a first chamber with afirst open end, a second open end, and a lumen extending therebetween, asupplemental chamber having a first open end, a second open end, and alumen therebetween, and a stopper positioned between the second end ofthe first chamber and the second end of the supplemental chamber. In oneembodiment, the stopper is elastomeric, self-lubricating, encloses amagnetic bar or one or more magnetic particles, or is a one way-valve.

The stopper has two positions: a first position and a second position.The stopper is sealingly positioned between the second end of the firstchamber and the second end of the supplemental chamber in the firstposition. In this position, the stopper seals the contents of the firstchamber from the contents of the supplemental chamber. For example, thecircumference of the stopper contacts the inner wall of the container toprevent leakage of contents between the first and supplemental chamberin the first position. The stopper is in the second position after thestopper is displaced or compromised such that the stopper no longerseals the first chamber from the contents of the second chamber. Forexample, in the second position, the stopper is displaced, i.e.,unseated such that the circumference of the stopper is no longer incontact with the inner wall of the container. In another embodiment ofthe stopper, e.g., a one-way valve, the stopper is compromised when thevalve opens to permit the contents of the first chamber and the contentsof the supplemental chamber to come in contact with one another.

In the second position of the stopper, the lumen of the supplementalchamber is co-extensive and in fluid communication with the lumen of thefirst chamber.

In one embodiment, the reagent mixing container further includes aperforatable, self-sealing cap positioned at the first open end of thefirst chamber, a plunger sealingly positioned at the first open end ofthe supplemental chamber and slideably moveable in the lumen of thesupplemental chamber from the supplemental chamber first end towards thesupplemental chamber second end.

The reagent mixing container contains a lyophilized component of areagent and/or a liquid component of a reagent in at least one of thefirst chamber and the supplemental chamber. In one embodiment, forexample, the lyophilized reagent is a lyophilized PT (prothrombin time)reagent, lyophilized Thrombin Time reagent, or lyophilized bovinethrombin reagent. Alternatively, the liquid reagent is deionized water,a buffer, or concentrated reagents such as but not limited toconcentrated PT (prothrombin time) reagent, alternatively, aconcentrated D-Dimer reagent, or latex reagent.

In another aspect, the invention described herein is related to a systemfor automated mixing of reagents. The system includes the features ofthe dual chamber reagent mixing container described above. Also includedin the system is a clinical analyzer in one embodiment having anactuator for driving the plunger into the lumen of the supplementalchamber, a stepper motor for driving the actuator, an electro-magneticcoil (or magnet) for driving rotation of the reagent mixing container oractuating movement of the magnetic mixing bar or the magnetic beads.

In another aspect, the invention described herein is related to a methodfor automated reagent mixing. The method comprises a number of steps. Inone embodiment of the method, the dual chamber reagent mixing vialcomprising a first reagent component in the first chamber and a secondreagent component in the supplemental chamber is provided in the mixingsystem described above. The plunger is transferred from a non-activatedposition where it is positioned in the first end of the supplementalchamber to seal the opening at the first end, to an activated positionby advancing the plunger automatically in the lumen of the supplementalchamber towards the second end of the supplemental chamber. By advancingthe plunger towards the second end of the supplemental chamber, thecontents of the supplemental chamber, liquid, powder or gas, iscompressed against the stopper 16. In one embodiment, the stopper isdisplaced by compression of the contents of the supplemental chamberagainst the stopper to unseat the stopper. Alternatively, compression ofthe contents of the supplemental chamber against the one-way valvestopper compromises the stopper by opening the stopper. Followingunseating of the stopper, or opening of the one-way valve stopper, thelumen of the first chamber and the lumen of the supplemental chamber arein fluid communication.

In one embodiment of this aspect of the invention, after the stopper isdisplaced or compromised, the first reagent and the second reagentcomponents are mixed together by actuating a rotor to rotate the reagentmixing container. In an alternative embodiment, the components of thereagents are mixed by rotation of the stopper encasing a magnetic bar orby rotation of at least one magnetic particle.

In yet another aspect, the invention described herein is related to amethod for introducing reagent components into the dual chamber reagentmixing container described above. The reagent container described aboveis readied with the perforatable sealing cap unsealed and the plunger inits sealing position in the opening at the first end of the supplementalchamber. A liquid form of the second component is lyophilized in thelumen of the supplemental chamber while the sealing cap remainsunsealed. The stopper is securely positioned at the interface of thesecond end of the first chamber and the second end of the second chamberto seal the contents of the lumen of the first and second chambers fromeach other. A liquid is introduced into the lumen of the first chamberand the sealing cap is pushed into the opening at the first end of thefirst chamber to form a seal whereby the reagent mixing container isrendered air tight.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 illustrates a perspective view of the reagent mixing containeraccording to an embodiment of the invention;

FIG. 2A illustrates a transverse section of the reagent mixing containerillustrated in FIG. 1;

FIG. 2B illustrates a transverse section of the reagent containerillustrated in FIG. 2A with the plunger activated to displace thestopper into a second position thereby releasing the contents of thesupplemental chamber into the first chamber;

FIG. 3 illustrates a transverse section of a magnetic bar/stoppercombination according to an embodiment of the invention;

FIGS. 4A and 4B illustrate a transverse section of a one-way valvestopper according to another embodiment of the invention;

FIG. 5A illustrates a schematic view of a system for automated mixing ofreagents in a clinical analyzer including the reagent mixing containershown in a non-activated transverse section, a linear actuator and astepper motor, and an electromagnetic coil according to an embodiment ofthe invention;

FIG. 5B illustrates a schematic view of the reagent mixing container ofFIG. 5A with the plunger partially activated and the magneticbar/stopper combination released into the lumen of the first chamber;

FIG. 5C illustrates a schematic view of the reagent mixing container ofFIGS. 5A and 5B with the plunger fully activated, the reagent in thelumen of the second chamber moved into the lumen of the first chamberand mixed by rotating the magnetic bar/stopper combination or byrotating the reagent mixing container;

FIG. 6A illustrates a first step in preparing a lyophilized reagent inthe first chamber according to one embodiment of the invention;

FIG. 6B illustrates a second step of the lyophilization procedureillustrated in FIG. 6A according to one embodiment of the invention.

DESCRIPTION OF THE INVENTION

Described below is an automated dual chamber reagent mixing containerfor separately storing and automatically mixing together at least twostored reagents, methods for mixing stored reagents, and methods formanufacturing the dual chamber reagent mixing container with its contentfor use in an automated clinical analyzer, including hemostasisanalyzers, immunoassay analyzers, and chemistry analyzers, to name afew. Various combinations and arrangements of reagents and chambers arecontemplated by the invention.

These and other objects, along with advantages and features of thepresent invention described herein, will become apparent throughreferences to the following description and the claims. Furthermore, itis to be understood that the features of the various embodimentsdescribed herein are not mutually exclusive and can exist in variouscombinations and permutations.

In one aspect, the invention is directed to a device for storage of andautomated mixing of two reagents for use in an automated clinicalanalyzer, for example, Hemostasis Analyzer, ACLTOP® 300, 500 and 700series, (Instrumentation Laboratory® Company, Bedford, Mass.).

Referring to FIGS. 1 and 2A, a reagent mixing container 10 according tothe invention includes a first chamber 12, a supplemental chamber 14, astopper 16, a plunger 18, a resealable cap 20, and, optionally, aprotective cap 24. The first chamber 12 has a first end 11 positioned onthe top of the container 10, a second end 13 opposite the first end 11positioned along the long axis of the container 10 below the first end11, and a lumen 26 extending between the first end 11 and the second end13 of the first chamber 12. The volume of the first chamber may be inthe range of about 10 ml to 5,000 ml, 10 ml to 1000 ml, 10 ml to 500 ml,10 ml to 100 ml, 100 ml to 500 ml, 100 ml to 250 ml, or 100 ml.

The supplemental chamber 14 has a second end 15 positioned adjacent thesecond end 13 of the first chamber 12, a first end 17 opposite thesecond end 15, and a lumen 28 extending between the first end 17 and thesecond end 15 of the supplemental chamber 14. The volume of thesupplemental chamber 14 may be about 0.1 ml to 100 ml. Each of the firstchamber 12 and the supplemental chamber 14 may enclose one or moreliquid or dry reagents.

With continued reference to FIG. 1, the plunger 18, has two positions: asealing or first position in which the plunger 18 seals the first end 17of the supplemental chamber 14 (shown in FIG. 1) forming a lumen 28 inthe supplemental chamber 14, and an actuated or second position in whichthe plunger 18 is actuated by an external force, described in greaterdetail below with respect to FIG. 2B and illustrated in FIGS. 5A-5C, tomove the plunger in the lumen 28 from the first end 17 towards thesecond end 15 of the supplemental chamber 14 as indicated by the arrowin FIG. 2B. When the plunger 18 is moved as indicated by arrow, thestopper 16 is displaced from circumferential contact with the inner wall11 of the container 10.

The resealable cap 20 is positioned at an opening of the first end ortop 11 of the first chamber 12 and is perforatable when a probe, such asa pipette or needle (not shown) pierces the cap 20. The cap 20 resealswhen the probe is withdrawn. When the reagent mixing container 10 is notin use, the cap 20 is closed to seal the first chamber 12 from room airthereby preventing evaporation of contents within the chambers of thereagent mixing container 10.

Referring to FIG. 2B, the reagent mixing container 10 has a stopper 16,positioned at the interface between the second end 13 of the firstchamber 12 and the second end 15 of the supplemental chamber 14separating the contents, such as a reagent component of the firstchamber 12 (represented by stippling) from the contents, such as anotherreagent component, of the supplemental chamber 14. The stopper 16 issecured in position in contact with the inner wall 11 of the container10 and seals the lumen 26 of the first chamber 12 from the contents ofthe lumen 28 of the supplemental chamber 14 while the reagent mixingcontainer 10 is stored until the stopper 16 is compromised or displaced.The stopper 16 is displaced or compromised by displacing the stopperfrom contact with the inner wall 11 or the stopper 16 is opened(compromised) by opening a one-way valve stopper positioned between thefirst and supplemental chambers. By displacement, the stopper 16 isfreed from its seated position against the inner wall of the container10 and is able to freely move within the container lumen.

When the stopper 16 is displaced or the stopper 16 comprises a one wayvalve that is opened, the lumen 28 of the supplemental chamber 14, andthe lumen 26 of the first chamber 12, are co-extensive and in fluidcommunication.

In various embodiments of the invention, the stopper 16 is elastomeric,for example, rubber, or plastic, and/or coated with a self-lubricatingmaterial such as Teflon® (polytetrafluoroethylene). In one embodiment ofthe invention, the stopper 16 is a compressible material, such as rubberor plastic, or a self-lubricating coated material 23 any one of whichencloses a rotatable magnetic bar 22, illustrated in FIG. 3 or,alternatively encloses one or more magnetic particles (not shown). Inone embodiment of the invention, the stopper 16 is a one way valvedescribed below with reference to FIGS. 4A-4B.

In an embodiment of the invention, discussed in greater detail withrespect to FIGS. 5A-5C below, the stopper 16 has a first position and asecond position. In a first position the stopper 16 is removablypositioned at the interface between the second end 13 of the firstchamber 12 and the second end 15 of the supplemental chamber 14 with thecircumference of the stopper 16 in contact with the inner wall 11 of thecontainer 10. In this position, the stopper 16 seals the reagentcontents of the first chamber 12 from the reagent contents of thesupplemental chamber 14 as long as the stopper 16 remains undisruptedand the integrity of the stopper 16 is uncompromised. In thisembodiment, while the stopper 16 is in the first position, the contentsin each of the two chambers of the reagent container 10 remain separatedand can be stored in this way for periods of time extending to days,weeks, months or even one or more years, prior to use withoutevaporation or loss of reagent activity.

As illustrated in FIG. 3, in one embodiment of the invention the stopper16 encases a magnetic bar 22 by a material such as but not limited torubber, a polymer such as plastic, or Teflon®. The stopper 16 isreleasably secured at the interface of the lumen 26 at the second end 13of the first chamber 12 and the lumen 28 at the second end 15 of thesecond chamber 14. In this embodiment, the stopper 16 is secured bycompression of the stopper 16 against the interior surface of the wallsof the container 10. In one embodiment, the stopper 16 has a diameterslightly larger than the diameter of the aforementioned interface sothat it is secured by friction but releasable into the lumen of thecontainer. When a force is applied to the stopper, for example, duringcompression of the contents of the supplemental chamber or when itsintegrity is compromised (such as opening a one-way valve) the stoppermoves to a second position which permits the contents of the firstchamber to reach the contents of the supplemental chamber.

Referring to FIGS. 4A and 4B, in another embodiment of the automatedreagent mixing container 10 of the invention, the stopper 16 is aone-way valve. As illustrated in FIG. 4A, the one-way valve stopper 16in the closed (first) position prevents mixing of the contents of thelumen 26 of the first chamber 12 with the contents of the lumen 28 ofthe supplemental chamber 14. When positive pressure is applied by theplunger 18 while it is advanced towards the second end 15 in the lumen28 of the supplemental chamber 14 as described below, the one-way valve16 is compromised and opens into the lumen 26 of the first chamber 12 asillustrated in FIG. 4B. The compromised valve/stopper 16 permits thecontents of the lumen 28 of the supplemental chamber 14 to reach thecontents in the lumen 26 of the first chamber 12.

As illustrated in FIGS. 4A and 4B, in a particular embodiment one ormore magnetic beads 30 may be positioned in the lumen 28 of thesupplemental chamber 14 or positioned within the one-way valve 16. Whenthe one-way valve/stopper is compromised and opens under pressureapplied by the plunger 18 as described above, the one or more magneticbeads 30 are forced into the lumen 26 of the first chamber 12 andfacilitate mixing of the components of the first chamber 12 andsupplemental chamber 14 when a magnetic field is applied to the magneticbead 30. The magnetic field may be generated by an electromagnetic coil36 or a magnet as shown in FIG. 5C.

In another aspect, the invention is directed to a system for automatedmixing of reagents in a clinical analyzer.

Referring now to FIG. 5A, the system according to the invention includesthe reagent mixing container 10 described above, an actuator 32, such asa linear actuator, a motor 34, such as a stepper motor, and optionallyan electromagnetic coil or magnet 36 for generating a magnetic field.The actuator 32 is operably positioned to slideably advance the plunger18 in the lumen 28 from the first end 17 of the supplemental chamber 14towards the second end 15 of the supplemental chamber 14 up to and,optionally, slightly beyond the stopper 16 of the reagent mixingcontainer 10.

In the configuration represented by FIGS. 5B and 5C, the reagent mixingcontainer 10 performs a mixing function. Referring to FIGS. 5B and 5C,the components, such as liquid or dry components of a reagent, inchambers 12 and 14 of the container 10, are combined when the stopper 16is displaced, for example, by unseating the stopper 16 from itsremovably secured first position at the interface of the second end 13of the first chamber 12 and the second end 15 of the second chamber 14to the second position. In the second position, illustrated in FIG. 5B,the rubber stopper 16 is unseated and the contents of the first chamber12 come in contact with the contents of the supplemental chamber 14.Alternatively, as described above and illustrated in FIGS. 4A and 4B,the components in first chamber 12 contacts the components ofsupplemental chamber 14 when the one-way valve-stopper 16 translatesfrom a closed to an open position. Following the opening of the valveand contact of the components of the two chambers, mixing of thecomponents can take place.

According to one embodiment of the method of the invention, the stopper16 is unseated or the valve-stopper 16 is opened by the action of theplunger 18 when the plunger 18 is advanced by, for example, the linearactuator 32 activated by the stepper motor 34, towards the stopper 16 inthe lumen 28 of the supplemental chamber 14 from the first end 17 of thesupplemental chamber 14 towards the second end. Advancing the plunger18, as illustrated in FIGS. 5B-5C, compresses the contents of the lumen28 of the supplemental chamber 14 whereby the compressed contents pushon the stopper 16 disrupting the stopper 16 from its first positionillustrated in FIG. 5A to the second position illustrated in FIGS. 5B-5Cor opens the valve-stopper 16 to a second position as illustrated inFIG. 4B. The reagent component contents (liquid or powder) in the lumen28 of the supplemental chamber 14 are advanced into the lumen 26 of thefirst chamber 12. Following introduction of the reagent componentcontents of the supplemental chamber 14 into the lumen 26 of the firstchamber 12, the reagent components of the first chamber 12 contact thereagent components of the second chamber 14.

Mixing of the reagents components of the first chamber 12 with thereagents components of the second chamber 14 begins after the stopper 16is in the second position. As discussed above, in one embodimentaccording to the method of the invention, the stopper encasing amagnetic stir bar 22 or alternatively one or more magnetic particles(not shown) is activated. In an embodiment of the method of theinvention, the magnetic mixing bar 22 or the one or more magneticparticles is operably joined to the electromagnetic coil or a magnet 36.The electromagnetic coil 36 actuates the magnetic bar 22 or magneticparticles, illustrated in FIG. 5C, i.e., causing the magnetic bar 22 torotate or the magnetic particles to move inside the lumen 26 of thefirst chamber 12 in the reagent mixing container 10, to obtain ahomogeneous mixing of the reagent components of the first chamber 12 andthe supplemental chamber 14.

Alternatively, the regent components of the first chamber 12 and thesupplemental chamber 14 are mixed together in the first chamber 12 byrotation, such as by oscillation, of the reagent mixing container 10.

In yet another aspect, the invention is directed to a method for makingthe reagent mixing container 10 described above. Referring to FIG. 6A,in one embodiment of the invention, a method for preparing a lyophilizedreagent in the first chamber 12 is shown. In this embodiment, theplunger 18 is positioned to seal the first end 17 of the supplementalchamber 14, a diluent is added to the lumen 28 of the supplementalchamber 14, followed by securely positioning the stopper 16 at thesecond end 13 of the first chamber 12 to seal the lumen 28 of thesupplemental chamber 14. A homogenous or a heterogeneous liquid reagentcomprising two or more components is added to the lumen 26 of the firstchamber 12. The liquid reagent may be, for example, a component of PTReadiPlasTin® (Instrumentation Laboratory Company, Bedford, Mass.), areagent used for coagulation testing. The first chamber 12 is partiallyfilled or completely filled with the liquid reagent to the top of thelumen 26, i.e., approaching the first end 11 of the first chamber 12.

Still referring to FIG. 6A, with the liquid reagent filled to a desiredlevel in the lumen 26 of the first chamber 12, the resealable cap 20,including a rubber septum 65, is kept in an open position. The openposition, as used herein, refers to loosely placing the resealable cap20 at the first end 11 of the chamber 12 without sealing the lumen 26 ofthe first chamber 12 from room air. For example, the resealable cap 20includes a rubber septum 65 that has openings (vents indicated asarrows) at contact points between the rubber septum 65 and the openingat the first end 11 of the first chamber 12. The openings permit vacuumventing of the pre-filled liquid reagent in the lumen 26 of the firstchamber 12. Vacuum venting of the liquid reagent in the lumen 26 iscarried out by the application of negative pressure with the use of apressure regulator (not shown) to control the vent-up rate. Vacuumventing proceeds until the components of the pre-filled liquid reagentin the first chamber 12 are lyophilized. The duration and pressure forvacuum venting of the liquid reagent may vary depending on the volumeand/or the density of the liquid in the lumen 26.

Referring now to FIG. 6B, at the end of lyophilization, i.e., once theliquid reagent is lyophilized in the lumen 26 of the first chamber 12 togenerate a lyophilized product, the resealable cap 20 is brought to aclosed position from the previously open position. To bring theresealable cap 20 to the closed position, the resealable cap 20 ispushed towards the lumen 26 (as indicated by the arrow), at the firstend 11 of the chamber 12 to seal the chamber 12. Sealing is achievedwhen the rubber septum 65 attached to the resealable cap fits securelyinto the opening of the upper end 11 of the lumen 26 of the firstchamber 12. Sealing by the resealable cap at the first end 11 of thefirst chamber 12 prevents contact of the lyophilized reagents in thelumen 26 of the first chamber 12 with room air outside the dual chamberreagent mixing container 10. Sealing by the stopper at the second end 15of the supplemental chamber 14 prevents mixing of the contents of thefirst chamber 12 and the supplemental chamber 14 until the stopper 16 isdislodged or its integrity compromised (e.g., the integrity of a one-wayvalve stopper is moved from a closed to an open valve) to permit contactbetween the contents of the two chambers.

Referring to FIGS. 6A and 6B, vacuum venting of the liquid reagent inthe chamber 12 after lyophilization of the liquid reagent is done in amanner such that the outer brim 42 at the bottom of the reagent mixingcontainer 10 remains rested on a base 51 of a support unit 55. The base51 permits the plunger 18 to stay rested, i.e., undisturbed duringlyophilization of the liquid as well as during the sealing of thecontainer 10 once the liquid is lyophilized. The base 51 also preventsdiluents, if present in the supplemental chamber 14, to remainundisturbed.

Thus, in an embodiment of the invention, during or after lyophilizationof the liquid reagent in the reagent mixing container 10, the liquidreagent or the lyophilized version of the liquid reagent remainsseparated from the contents, such as a diluent or a concentrate that isplaced in the lumen 28 of the supplemental chamber 14. Upon sealing, thelyophilized powder is stored without loss of activity or evaporationuntil mixing of the lyophilized powder with a diluent is desired.

The advantages of the invention are that the reagent preparation, i.e.,lyophilization of a liquid reagent in the first chamber 12, and thereconstitution of the lyophilized component with a dilution solution,such as a diluent stored in the supplemental chamber 14, and mixing ofthe two reagent components can be accomplished automatically in aclinical analyzer without manual intervention, thereby eliminating thepossibility of erroneous reagent preparation due to human error,inadvertent contamination of an operator while mixing reagents manually,or inadvertent loss of reagents. In addition, because the volume of eachreagent components is defined by the size of the chambers 12 and 14 inthe container 10, the ratio at which the two components can be mixed canbe controlled.

Exemplifications

An Exemplary Dual Chamber Reagent Mixing Container for HemostasisTesting in an Automated Clinical Hemostasis Analyzer Instrument

A specific non-limiting example of the dual chamber reagent mixingcontainer according to the invention is a reagent container forprothrombin time (PT) testing by a clinical hemostasis analyzerinstrument. The reagent for PT testing contains two components: adiluent and a concentrated PT reagent. The volume ratio between thediluent and the PT reagent is 19:1. Each PT test requires 100 μl dilutedPT reagent. A container useful for PT tests in an automated clinicalanalyzer would require between 500 to 1000 PT tests. A 1000 PT testcontainer requires 100 ml of diluted PT reagent, namely 95 ml diluentand 5 ml concentrated PT reagent. In the exemplary container shown inFIG. 1, the first chamber 12 would have a volume capacity greater than100 ml and initially contains 95 ml of diluent. The supplemental 14chamber contains 5 ml concentrated PT reagent. Upon actuation by theplunger 18, the 5 ml of concentrated PT reagent is moved into the lumen26 of the first chamber 72 following disruption or loss of integrity ofthe stopper 16. Actuators for moving the plunger 18 are integrated intothe hemostasis analytical instrument. An exemplary actuator 32 isillustrated in FIGS. 5A-5C, a stepper motor 34 is used to drive theplunger 18, and an electro-magnetic coil 36 is used to drive therotation of the magnetic bar-stopper 16 combination or at least onemagnetic bead. A self-contained PT reagent mixing container 10 withautomated reagent preparation, sustainable for 1000 PT assays, isachieved.

An Exemplary Dual Chamber Reagent Mixing Container for Analyte Testingin an Automated Clinical Analyzer

A second non-limiting example of the dual chamber reagent mixingcontainer according to the invention is a reagent container for latexreagent of D-Dimer testing. The latex reagent for D-Dimer is prepared bydiluting a concentrated latex reagent (Instrumentation LaboratoryCompany) with deionized water in a 1:1 volume ratio of concentratedlatex reagent:deionized water. Each D-dimer test requires 100 μl ofdiluted D-dimer latex reagent. A container for 1000 tests requires 100ml diluted-dimer latex reagent. Therefore, in this exemplary containeraccording to the invention, illustrated in FIG. 1, the first chamber 12would have a volume capacity larger than 100 ml and initially contains50 ml of deionized water, and the supplemental chamber 14 contains 50 mlconcentrated D-dimer latex reagent. Upon actuation by the plunger 18,the 50 ml of concentrated D-dimer latex reagent moves into the lumen ofthe first chamber 12 following disruption or loss of integrity of thestopper 16. Actuators for moving the plunger 18 are integrated into theanalytical instrument. An exemplary actuator 32 is illustrated in FIGS.5A-5C, a stepper motor 34 is used to drive the plunger 18, and anelectro-magnetic coil 36 is used to drive the rotation of the magneticbar/stopper 16 combination or at least one magnetic bead. Aself-contained D-dimer reagent mixing container 10 with automatedreagent preparation, sustainable for 1000 D-dimer assays, is achieved.

We claim:
 1. A method for automated reagent mixing in a clinicalanalyzer, comprising: providing a system for automated mixing ofreagents, comprising: a) a reagent mixing container comprising, (i) afirst chamber comprising a first end, a second end, and a lumenextending therebetween, (ii) a supplemental chamber comprising a firstend, a second end, and a lumen therebetween, wherein said second end ofsaid first chamber and said second end of said supplemental chamber areadjacent, (iii) a stopper comprising a first position and a secondposition, wherein in said first position said stopper is positioned toseal the lumen of said first chamber at said second end of said firstchamber and the lumen of said supplemental chamber at said second end ofsaid supplemental chamber, and in said second position, said stopper isdisplaced or compromised from said first position, whereby the lumen ofsaid supplemental chamber is co-extensive with the lumen of said firstchamber, and, (iv) a plunger sealingly positioned at the first end ofsaid supplemental chamber, and slideably moveable in the lumen of saidsupplemental chamber from the supplemental chamber first end towardssaid supplemental chamber second end; b) an actuator for driving theplunger into the lumen of the supplemental chamber, c) a stepper motorfor driving the actuator, and, d) an electro-magnetic coil for drivingrotation of the reagent mixing container; providing a first reagentcomponent in said first chamber and a second reagent component in saidsupplemental chamber; transferring the plunger by said actuator from anon-activated position at the first end of said supplemental chamber toan activated position at the second end of said supplemental chamber;releasing said stopper; rotating said reagent mixing container; andmixing said first reagent components and second reagents together. 2.The method of claim 1 wherein releasing said stopper comprisesdisplacing said stopper.
 3. The method of claim 1 wherein releasing saidstopper comprises opening a valve.
 4. A method for automated reagentmixing in a clinical analyzer, comprising: providing a system forautomated mixing of reagents, comprising: a) a reagent mixing containercomprising, (i) a first chamber having a first end, a second end, and alumen extending therebetween, (ii) a supplemental chamber having a firstend, a second end, and a lumen therebetween, wherein said second end ofsaid first chamber and said second end of said supplemental chamber areadjacent, (iii) a stopper comprising a first position and a secondposition, wherein in said first position said stopper is positioned toseal the lumen of said first chamber at said second end of said firstchamber and the lumen of said supplemental chamber at said second end ofsaid supplemental chamber, and in said second position, said stopper isdisplaced or compromised from said first position, whereby the lumen ofsaid supplemental chamber is co-extensive with the lumen of said firstchamber, (iv) a plunger sealingly positioned at the first end of saidsupplemental chamber, and slideably moveable in the lumen of saidsupplemental chamber from the supplemental chamber first end towardssaid supplemental chamber second end; b) an actuator for driving theplunger into the lumen of the supplemental chamber, c) a stepper motorfor driving the actuator; transferring the plunger by said actuator froma non-activated position to an activated position; releasing saidstopper; providing a first reagent component in said first chamber and asecond reagent component in said supplemental chamber; actuating a motorto rotate a magnetic bar, or at least one magnetic particle; and mixingsaid first reagent components and said second reagents together.
 5. Themethod of claim 4 wherein releasing said stopper comprises opening avalve.
 6. The method of claim 4 wherein releasing said stopper comprisesdisplacing said stopper.
 7. A method for introducing reagents into anautomatic reagent mixing container, comprising: providing a reagentmixing container comprising: (i) a first chamber comprising a first end,a second end, and a lumen extending therebetween, (ii) a supplementalchamber comprising a first end, a second end, and a lumen therebetween,wherein said second end of said first chamber and said second end ofsaid supplemental chamber are adjacent, (iii) a stopper, (iv) aperforatable, self-sealing cap positioned at the first open end of saidfirst chamber, and (v) a plunger sealingly positioned at the first endof said supplemental chamber, and slideably moveable in the lumen ofsaid supplemental chamber from the supplemental chamber first endtowards said supplemental chamber second end; sealing the first end ofsaid supplemental chamber with said plunger; introducing a liquidcomponent into said supplemental chamber; lyophilizing the liquidcomponent in said supplemental chamber; sealing said second end of saidsupplemental chamber with said stopper; introducing a liquid into saidfirst chamber; inserting said resealable cap into said first end of saidfirst chamber to form a seal, whereby said supplemental chamber and saidfirst chamber are air tight.